Multiple system atrophy prions retain strain specificity after serial propagation in two different Tg( SNCA *A53T) mouse lines

Previously, we reported that intracranial inoculation of brain homogenate from multiple system atrophy (MSA) patient samples produces neurological disease in the transgenic (Tg) mouse model TgM83 +/− , which uses the prion protein promoter to express human α-synuclein harboring the A53T mutation found in familial Parkinson’s disease (PD). In our studies, we inoculated MSA and control patient samples into Tg mice constructed using a P1 artificial chromosome to express wild-type (WT), A30P, and A53T human α-synuclein on a mouse α-synuclein knockout background [Tg( SNCA +/+ )Nbm, Tg( SNCA *A30P +/+ )Nbm, and Tg( SNCA *A53T +/+ )Nbm]. In contrast to studies using TgM83 +/− mice, motor deficits were not observed by 330–400 days in any of the Tg( SNCA )Nbm mice after inoculation with MSA brain homogenates. However, using a cell-based bioassay to measure α-synuclein prions, we found brain homogenates from Tg( SNCA *A53T +/+ )Nbm mice inoculated with MSA patient samples contained α-synuclein prions, whereas control mice did not. Moreover, these α-synuclein aggregates retained the biological and biochemical characteristics of the α-synuclein prions in MSA patient samples. Intriguingly, Tg( SNCA *A53T +/+ )Nbm mice developed α-synuclein pathology in neurons and astrocytes throughout the limbic system. This finding is in contrast to MSA-inoculated TgM83 +/− mice, which develop exclusively neuronal α-synuclein aggregates in the hindbrain that cause motor deficits with advanced disease. In a crossover experiment, we inoculated TgM83 +/− mice with brain homogenate from two MSA patient samples or one control sample first inoculated, or passaged, in Tg( SNCA *A53T +/+ )Nbm animals. Additionally, we performed the reverse experiment by inoculating Tg( SNCA *A53T +/+ )Nbm mice with brain homogenate from the same two MSA samples and one control sample first passaged in TgM83 +/− animals. The TgM83 +/− mice inoculated with mouse-passaged MSA developed motor dysfunction and α-synuclein prions, whereas the mouse-passaged control sample had no effect. Similarly, the mouse-passaged MSA samples induced α-synuclein prion formation in Tg( SNCA *A53T +/+ )Nbm mice, but the mouse-passaged control sample did not. The confirmed transmission of α-synuclein prions to a second synucleinopathy model and the ability to propagate prions between two distinct mouse lines while retaining strain-specific properties provides compelling evidence that MSA is a prion disease.